FIG 2 .

Effects of osmolarity on P. aeruginosa-induced blebs in human corneal epithelial cells. (a) Schematic of the experimental outline for infection prior to treatment with hyperosmotic media. Four hours post-inoculation, after blebs had formed, hyperosmotic media was added to the cells for an additional 3 h (using mannitol to achieve a final osmolarity of 400, 500, or 600 mOsM). (b) Schematic showing a similar experimental outline for induction of apoptosis without infection prior to osmotic treatment. (c) Representative images showing the effects of the hyperosmotic media (600 mOsM) on blebbing (see arrows). (d) Blebbing frequency in human corneal epithelial cells following infection in iso-osmolar media (15% ± 4%) compared to that in increasingly hyperosmolar media (2% ± 1% in 600 mOsM). (e) Blebbing frequency following induction of apoptosis in iso-osmolar media (57% ± 4%) compared to that in increasingly hyperosmolar media (91% ± 1% in 600 mOsM), *, P < 0.05; **, P < 0.01; ***, P < 0.001 (ANOVA with Dunnett’s multiple-comparison post hoc analysis).